In vitro proliferation and culture method for duck circovirus

A duck circovirus, in vitro propagation technology, applied in the direction of virus/bacteriophage, virus, single-stranded DNA virus, etc., can solve the problems of increased incidence of pathogenicity, no progress in virus in vitro propagation, and no cultivation of DuCV. Titer-increasing effect

Active Publication Date: 2018-11-20
INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI
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Problems solved by technology

[0003] DuCV was discovered in 2003. Since the virus does not cause visible lesions to host cells in most cases, the in vitro propagation of the virus has not progressed. Up to now, there is no feasible method for in vitro propagation and cultivation of DuCV, which cannot be completed in the laboratory. Virus isolation and identification
Therefore, there are many difficulties in the research on its pathogenic mechanism and harm, and the development of vaccines, and the understanding is still very superficial.
However, the current epidemiological studies have shown that the disease is common in duck populations in my country. Drawing on the existing research results of other circoviruses, it is speculated that the existence of DuCV may increase the risk of duck plague, duck viral hepatitis, and duck infectious serosa. colibacillosis, duck pasteurellosis, Escherichia coli disease and other common duck diseases and aggravated its pathogenicity, so in today's rapid development of duck industry scale and quantity, it is urgent to propose a method of in vitro multiplication and cultivation of DuCV

Method used

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  • In vitro proliferation and culture method for duck circovirus
  • In vitro proliferation and culture method for duck circovirus
  • In vitro proliferation and culture method for duck circovirus

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Embodiment Construction

[0022] A kind of in vitro propagation and culture method of duck circovirus, specifically comprises the following steps:

[0023] 1. Blood collection and separation of peripheral blood lymphocytes

[0024] Using heparin anticoagulant tubes (Jiangsu Kangjian Medical Supplies Co., Ltd.), the blood of adult Muscovy ducks was collected, mixed up and down slightly, stored in an ice bath and sent to the laboratory. Under aseptic operation, mix duck blood and PBS at a ratio of 1:2, and take 1 volume of lymphocyte separation medium (Ficoll-PaqueTMPREMIUM) in a 50ml centrifuge tube. Slowly add the mixture of duck blood and PBS to the upper layer of the lymphocyte separation medium to form a good layer. Transfer the centrifuge tube to a centrifuge for centrifugation, set the speed at 6000rpm / min for 20 minutes, and set the speed up and speed down of the centrifuge to the minimum. After centrifugation, the peripheral blood lymphocyte layer and lymphocyte separation liquid layer were co...

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Abstract

The invention provides an in vitro proliferation and culture method for a duck circovirus. The method comprises the following steps: (1) collecting healthy adult Muscovy duck blood for isolated culture of PBMC to prepare a PBMC cell suspension; and (2) adding a disease material which is detected duck circovirus positive into the PBMC cell suspension to be cultured. The invention provides the method of culturing DuCV by in vitro proliferation. The virus titer after virus inoculating culture is improved greatly, isolation and identification of the virus can be finished in a lab, and relatively good help is provided for researching the nosogenesis and damage of the virus.

Description

technical field [0001] The invention relates to an in vitro propagation and culture method of duck circovirus. Background technique [0002] Duck circovirus (Duckcircovirus, DuCV) is a new member of the Circovirus genus in the Circoviridae family. It can cause immunosuppression in ducks and cause harm to the duck industry. The clinical symptoms of ducks infected with duck circovirus DuCV are mainly poor development and obvious feather dystrophy, especially in the back spine (feathers). It is manifested as heterophilic inflammatory exudation of vesicles and tissues surrounding the vesicles. The general pathology of internal organs shows the symptoms of co-infection of duck infectious serositis. Histopathological examination of the bursa of Fabricius shows that lymphocyte damage, Necrosis, histiocytosis (Hattermann et al., 2003). [0003] DuCV was discovered in 2003. Since the virus does not cause visible lesions to host cells in most cases, the in vitro propagation of the v...

Claims

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Application Information

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IPC IPC(8): C12N7/00C12N5/078
CPCC12N5/0634C12N7/00C12N2750/10051
Inventor 李兆龙黄喻傅光华丰志华陈江华
Owner INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI
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